ORIGINAL PAPER Association of cotton leaf curl Multan betasatellite and Ageratum conyzoides symptomless alphasatellite with tomato leaf curl New Delhi virus in Luffa cylindrica in Pakistan Ifrah Anwar 1,2 & Hanan Arif Bukhari 1,2 & Nazia Nahid 2 & Kamran Rashid 2 & Imran Amin 1 & Shabnum Shaheen 3 & Khadim Hussain 2 & Shahid Mansoor 1 Received: 31 May 2019 /Accepted: 29 October 2019 # Australasian Plant Pathology Society Inc. 2019 Abstract Luffa plants showing symptoms of begomovirus infection were collected from Vehari, Punjab in 2016. RCA and PCR ampli- fications of begomovirus genomic components showed the presence of a typical Old World bipartite begomovirus Tomato leaf curl New Delhi virus (ToLCNDV). Interestingly, Ageratum conyzoides symptomless alphasatellite and CLCuMB were also found associated with bipartite begomoviruses in the same host, however, no monopartite begomovirus was detected. To the best of our knowledge, this is the first report of ToLCNDV with associated satellites infecting L. cylindrica in Pakistan. The importance of begomovirus extended host range with different combinations of associated satellites has been discussed. Keywords Alphasatellite . Betasatellites . Geminivirus . Luffa cylindrica . Phylogenetic studies Introduction Begomoviruses belonging to family Geminiviridae largely known for causing various plant diseases across the world (Brown et al. 2012) and transmitted by insect vector whitefly (Bemisia tabaci). Begomoviruses constitute monopartite and bipartite genomic form having one and two circular single stranded DNA components respectively. In bipartite begomoviruses, genomic components are named as DNA-A and DNA-B (Harrison and Robinson 1999). Monopartite begomoviruses consist of a single genomic component about 2.8 kb in size. Their genome is homologous to DNA-A of bipartite begomoviruses. Most of the monopartite begomoviruses are usually associated with satellite DNA mol- ecules. The alphasatellite and betasatellites associated with monopartite viruses are single stranded DNA molecules near- ly 1400 bp in size (Briddon et al. 2001; Mansoor et al. 1999). Unlike monopartite begomoviruses, only a few bipartite begomoviruses have been found associated with betasatellites (Nawaz-ul-Rehman and Fauquet 2009). Recently, the pres- ence of some alphasatellites have also been reported with bi- partite begomoviruses (Jeske et al. 2014; Paprotka et al. 2010; Romay et al. 2010; Zhang et al. 2016). Tomato leaf curl New Delhi virus (ToLCNDV) is a bipartite begomovirus which has been reported from a large number of plants including members of families Malvaceae, Solanaceae and Cucurbitaceae (Haider et al. 2006; Hussain et al. 2004; Tahir and Haider 2005). In 2016 in Vehari district of Punjab Pakistan, there was devastating disease incidence with typical symptoms of vein yellowing, yellow spots on the leaf and leaf curling (Fig. 1). In the present study, we have found ToLCNDV in a novel host Luffa cylindrica from Pakistan. Recently, ToLCNDV along with alphasatellite and betasatellites have been reported from pumpkin (Anwar 2017). However to the best of our knowledge, this is the first report of ToLCNDV infecting luffa in Pakistan. Luffa (Luffa cylindrica; family Cucurbitaceae) known by several local names including vegetable sponge, sponge gourd * Khadim Hussain hussaink@gcuf.edu.pk * Shahid Mansoor shahidmansoor7@gmail.com 1 Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan 2 Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan 3 Department of Botany, Lahore College for Women University, Lahore, Pakistan Australasian Plant Pathology https://doi.org/10.1007/s13313-019-00668-6